Probing conserved helical modules of portal complexes by mass spectrometry-based hydrogen/deuterium exchange
Probing conserved helical modules of portal complexes by mass spectrometry-based hydrogen/deuterium exchange
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DOI：
10.1016/j.jmb.2008.03.004
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发表时间：
2008-09-05
影响因子：
5.6
通讯作者：
Prevelige, Peter E., Jr.
中科院分区：
文献类型：
--
作者：
Kang, Sebyung;Poliakov, Anton;Prevelige, Peter E., Jr.
双链 DNA 噬菌体 P22 具有由 84 kDa porta I 蛋白亚基组成的环形十二聚体复合物，形成噬菌体 DNA 包装马达的中央通道。 P22门静脉复合体的整体形态与Phi29、SPP1、T3、T7噬菌体和单纯疱疹病毒的门静脉复合体相似。 P22 门蛋白的二级结构预测及其在 Phi29 门蛋白复合物晶体结构上的穿线表明，P22 门蛋白复合物共享在 Phi29 门蛋白复合物的十二聚体界面中发现的保守螺旋模块。为了识别 P22 门环复合物中亚基间接触所涉及的氨基酸并验证线程模型，我们对 P22 和十二聚体 Phi29 门户的单体和体外组装的门户蛋白进行了比较氢/氘交换分析。氢/氘交换实验提供了 P22 门户复合体中亚基间相互作用的证据，类似于 Phi29 门户中的相互作用，映射到预测为保守螺旋模块的区域。 (C) 2008 Elsevier Ltd. 保留所有权利。
The Double-stranded DNA bacteriophage P22 has a ring-shaped dodecameric complex composed of the 84 kDa porta I protein subunit that forms the central channel of the phage DNA packaging motor. The overall morphology of the P22 portal complex is similar to that of the portal complexes of Phi29, SPP1, T3, T7 phages and herpes simplex virus. Secondary structure prediction of P22 portal protein and its threading onto the crystal structure of the Phi29 portal complexes suggested that the P22 portal protein complex shares conserved helical modules that were found in the dodecameric interfaces of the Phi29 portal complex. To identify the amino acids involved in intersubunit contacts in the P22 portal ring complexes and validate the threading model, we performed comparative hydrogen/deuterium exchange analysis of monomeric and in vitro assembled portal proteins of P22 and the dodecameric Phi29 portal. Hydrogen/deuterium exchange experiments provided evidence of intersubunit interactions in the P22 portal complex similar to those in the Phi29 portal that map to the regions predicted to be conserved helical modules. (C) 2008 Elsevier Ltd. All rights reserved.